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Method for preparing theanine by using biological method

A theanine and biological method technology, which is applied in the field of biological preparation of theanine, can solve the problems of complex enzyme powder preparation, unreported industrial production process route, complicated separation and purification operations, etc., and achieves simple composition and low cost of the reaction solution. , post-processing simple effects

Inactive Publication Date: 2015-04-29
NANTONG LICHENG BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The second is the method of chemical organic synthesis of theanine. The products of this method include D and L-type theanine isomers, which need to be disassembled to obtain the L-type product, which makes the separation and refining operation complicated.
(2) In 2008, a method for producing theanine using glutamyl transpeptidase from Bacillus subtilis was disclosed. This method is safe and reliable, but there are problems in the preparation of enzyme powder and the use of ethanol in the process. Patent No. CN 101343618
In the existing method, the process route of industrialized production has not been reported, especially the method for producing theanine by adding substrates in batches by Pseudomonas nitroreductors

Method used

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  • Method for preparing theanine by using biological method
  • Method for preparing theanine by using biological method

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Experimental program
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Embodiment 1

[0026] (1) Cell culture

[0027] After the nitroreductor Pseudomonas strains are activated, the first-level seed culture, the second-level seed culture, and finally enter the fermenter culture, the inoculation amount is 3% (V / V, seed liquid / fermentation liquid), 30 ° C, stirring The speed was controlled at 130rpm, and the ventilation rate was 20m3 / h. After 4 hours, the stirring speed and ventilation rate were adjusted to maintain the dissolved oxygen not lower than 60. After culturing for 18 hours, the bacteria were collected by centrifugation.

[0028] Seed medium raw materials (in g / L): glucose 15, yeast extract 1, sodium glutamate 10, MgSO4 7H2O 0.5, K2HPO4 0.5, KH2PO4 0.5, EDTA-Fe 0.1, defoamer 0.2, pH 7.3

[0029] Fermentation medium raw materials (in g / L): glucose 15, yeast extract 1, sodium glutamate 10, MgSO4 7H2O 0.5, K2HPO4 0.5, KH2PO4 0.5, EDTA-Fe 0.1, defoamer 0.2, pH 7.3

[0030] (2) Production of theanine

[0031] Theanine is produced by the transformation meth...

Embodiment 2

[0041] The method for making the theanine powder product is as follows: the theanine clear liquid after membrane filtration obtained in step (4) of Example 1 is processed as follows:

[0042] (1) Vacuum concentration: Concentrate using a three-effect plate evaporator. When the feed liquid is concentrated to a theanine content ≥ 80g / L, stop concentrating and collect the concentrate.

[0043] (2) Spray drying: add maltodextrin to the concentrated solution, adjust the solid content to 30%, and spray dry to obtain a theanine powder product with a mass content of 50%.

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Abstract

The invention discloses a method for preparing theanine by using a biological method. The method comprises the steps of bacteria preparation, cell transformation, decoloring, electrodialysis, preparation of theanine powder products or theanine crystal products, and the like. According to the invention, free pseudomonas nitroreducens is adopted for synthesizing theanine by catalysis, and by using a method for adding substrates in batches, the yield of theanine is improved, the cost is lowered, and a high-concentration reaction is realized. A technological enzyme reaction system does not contain culture elements such as a carbon source, a nitrogen source, and the like, so that compared with fermentation liquor, reaction liquid is simple in composition, therefore, the product purity can be greatly improved; and the method has the advantages of easiness for separating an enzyme source from the reaction liquid, simple post-processing, short production cycle, high yield, small environmental pollution, low cost, and the like.

Description

technical field [0001] The invention relates to a method for preparing theanine by a biological method. Background technique [0002] L-theanine, also known as N-ethyl-γ-glutamine, was first isolated from green tea in 1949 and is a unique amino acid in tea. Theanine does not participate in protein synthesis and exists in free form, accounting for 40% to 70% of the total free amino acids in tea. It is an effective taste substance in green tea, and its content directly determines the quality of tea. In addition, theanine also has the effects of lowering blood pressure, anti-tumor, protecting nerves, relaxing, anti-depression, anti-fatigue, improving immunity, and reducing body fat. L-theanine has a variety of physiological functions, good stability and safety, so it has been widely used in the fields of food and pharmaceutical development. [0003] There are three main methods of manufacturing theanine: one is the method of extracting from tea, which is limited by tea resour...

Claims

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Application Information

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IPC IPC(8): C12P13/04C12R1/38
Inventor 帅玉英孙怡吴晓花顾钦青沈宇峰
Owner NANTONG LICHENG BIOLOGICAL ENG
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